Diagnosis of human toxocariasis currently relies on serologic tests that use Toxocara excretory-secretory (TES) antigen to detect immunoglobulin G (IgG) antibodies to the larvae. In general, however, these assays do not have adequate specificity for use in countries in which other soil-transmitted helminths are endemic. The use of recombinant antigens in these assays, however, is promising for improving the specificity of the diagnosis of toxocariasis. Toward this goal, we developed an IgG4 enzyme-linked immunosorbent assay (ELISA) involving three recombinant antigens: rTES-30USM (previously produced), rTES-26, and rTES-120. The latter two antigens were produced by reverse transcription-PCR cloning; subcloned into glutathione S-transferase (GST)-tagged and His-tagged prokaryotic expression vectors, respectively; and expressed in Escherichia coli. The recombinant proteins were subsequently purified by affinity chromatography using GST and His-Trap resins. The diagnostic potential of each purified recombinant antigen was tested with various immunoglobulin classes (IgG, IgM, and IgE) and IgG subclasses. The IgG4 ELISA was determined to have the highest specificity and was further evaluated using a panel of serum samples. The rTES-26 IgG4 ELISA showed 80.0% (24/30 samples positive) sensitivity, and both the rTES-30USM IgG4 ELISA and rTES-120 IgG4 ELISA had 93.0% (28/30) sensitivity. Combined use of rTES-120 and rTES-30 IgG4 ELISA for the diagnosis of toxocariasis provided 100% sensitivity. The specificities of rTES-26, rTES-30USM, and rTES-120 antigens were 96.2%, 93.9%, and 92.0%, respectively. These results indicate that the development of a diagnostic test using the three recombinant antigens will allow for more-accurate detection of toxocariasis.Human toxocariasis is a worldwide parasitic zoonosis, caused most commonly by the intestinal parasites the dog roundworm (Toxocara canis) and also the cat roundworm (Toxocara cati) (2). It commonly manifests as visceral larva migrans, ocular larva migrans, and covert toxocariasis. Toxocariasis is probably one of the most common zoonotic helminthiases in temperate climates and developed countries (20). In Malaysia, the seroprevalence rate is approximately 20% (6).Definitive diagnosis of toxocariasis is based on the detection of Toxocara larvae from biopsy tissues, but this test is timeconsuming and difficult to perform. Therefore, diagnosis is commonly based on clinical and serologic diagnosis. Currently, common routine serology tests are applied for detection, such as commercial immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) kits in which Toxocara excretorysecretory (TES) antigens obtained from culture of T. canis second-stage (L2) larvae are used (22). The use of the native TES antigen for serodiagnosis of human toxocariasis is a laborious and time-consuming technique, and the production capacity is limited by the culture volume (22). Further, the specificity is often low, especially in developing countries, where infections with helminths that cause c...

The thrombopoietin receptor (MPL) has been shown to be mutated (MPL W515L) in myelofibrosis and thrombocytosis yet new approaches to treat this disorder are still required. We have previously shown that transcriptome and proteomic effects do not correlate well in oncogene-mediated leukemogenesis. We therefore investigated the effects of MPL W515L using proteomics. The consequences of MPL W515L expression on over 3300 nuclear and 3500 cytoplasmic proteins were assessed using relative quantification mass spectrometry. We demonstrate that MPL W515L expression markedly modulates the CXCL12/CXCR4/CD45 pathway associated with stem and progenitor cell chemotactic movement. We also demonstrated that MPL W515L expressing cells displayed increased chemokinesis which required the MPL W515L-mediated dysregulation of MYC expression via phosphorylation of the RNA transport protein THOC5 on tyrosine 225. In addition MPL W515L expression induced TGFβ secretion which is linked to sphingosine 1-phosphate production and the increased chemokinesis. These studies identify several pathways which offer potential targets for therapeutic intervention in the treatment of MPL W515L-driven malignancy. We validate our approach by showing that CD34+ cells from MPL W515L positive patients display increased chemokinesis and that treatment with a combination of MYC and sphingosine kinase inhibitors leads to the preferential killing of MPL W515L expressing cells.

Limitations in existing anticoagulants have prompted a search for the natural origin of anticoagulants. Since there is an increasing need to source for pharmacological and medicinal materials from a plant source, thus an exploratory effort towards identifying and characterizing new anticoagulants from plants is worthwhile. Clinacanthus nutans (C. nutans) commonly known as Sabah snake grass belongs to Acanthaceae family was chosen since it has therapeutic and medicinal benefits. This plant possessed anti-oxidant, anti-microbial, anti-inflammatory, and anti-diabetic activities. This study was carried out to identify the anticoagulant activities of C. nutans leaves in aqueous and methanol extracts at different concentrations based on prothrombin time (PT) and activated partial thromboplastin time (aPTT). The dried leaves of C. nutans were ground into a fine powder and extracted using aqueous and methanol. Anticoagulant assays of PT and aPTT were done on three different concentrations of C. nutans aqueous and methanol extracts (10 mg/mL, 20 mg/mL, and 30 mg/mL) and control group respectively. PT and aPTT of plasma samples were prolonged in both types of extracts (p<0.05). This study highlights that the anticoagulant activity of aqueous and methanol extracts of C.nutans affects the intrinsic and extrinsic pathway, possibly due to the inhibitory action of the clotting factors. These results suggested that aqueous and methanol extracts of C. nutans exhibited anticoagulant activity at certain concentrations.

Autism Spectrum Disorder (ASD) is a range of neurodevelopmental disorders predominantly characterized by impaired social functioning and communication disturbances. Symptoms of autistic children may include repetitive movements, intense focus on one item, severe tantrum or self-abusive like behavior. In Malaysia, autism is still not clear and the public seems to lack good information or experience with people with ASD. Therefore, this study was investigated to describe the level of knowledge and attitude on ASD among public in Malaysia. A cross sectional study was conducted and Google link questionnaires containing demographic data, level of knowledge and attitude was distributed online via social media. Of 385 respondents, only 296 that meet the inclusion and exclusion criteria. Collected data were analyzed using Microsoft Excel 2013 and IBM Statistical Package for the Social Sciences (SPSS) version 20. The findings suggested that public in Malaysia have a moderate level of knowledge (72%) and low level of positive attitude (57%) towards ASD. Even there are many Malaysian people have heard about autism but still many of them do not really understand the nature of children with autism. Some of them even cannot recognize the physical appearance of autistic children. We can conclude that lack of knowledge can lead to the negative attitudes towards autistic individuals and families. Further awareness strategy and giving the right information of the autism via social media and educational campaign can decrease the discrimination and stigmatization on ASD among public.